Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

Click here for more information

Sign In to gain access to subscriptions and/or personal tools.
Journal of Biomolecular Screening
This Article
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
1087057109343207v1
14/8/956    most recent
Right arrow References
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to Saved Citations
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via Scopus
Google Scholar
Right arrow Articles by Antczak, C.
Right arrow Articles by Djaballah, H.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Antczak, C.
Right arrow Articles by Djaballah, H.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

Live-Cell Imaging of Caspase Activation for High-Content Screening

Christophe Antczak

High Throughput Screening Core Facility, Memorial Sloan-Kettering Cancer Center, New York

Toshimitsu Takagi

High Throughput Screening Core Facility, Memorial Sloan-Kettering Cancer Center, New York

Christina N. Ramirez

High Throughput Screening Core Facility, Memorial Sloan-Kettering Cancer Center, New York

Constantin Radu

High Throughput Screening Core Facility, Memorial Sloan-Kettering Cancer Center, New York

Hakim Djaballah

High Throughput Screening Core Facility, Memorial Sloan-Kettering Cancer Center, New York, djaballh{at}mskcc.org

Caspases are central to the execution of programmed cell death, and their activation constitutes the biochemical hallmark of apoptosis. In this article, the authors report the successful adaptation of a high-content assay method using the DEVDNucView488TM fluorogenic substrate, and for the first time, they show caspase activation in live cells induced by either drugs or siRNA. The fluorogenic substrate was found to be nontoxic over an exposure period of several days, during which the authors demonstrate automated imaging and quantification of caspase activation of the same cell population as a function of time. Overexpression of the antiapoptotic protein Bcl-XL, alone or in combination with the inhibitor Z-VAD-FMK, attenuated caspase activation in HeLa cells exposed to doxorubicin, etoposide, or cell death siRNA. This method was further validated against 2 well-characterized NSCLC cell lines reported to be sensitive (H3255) or refractory (H2030) to erlotinib, where the authors show a differential time-dependent activation was observed for H3255 and no significant changes in H2030, consistent with their respective chemosensitivity profile. In summary, the results demonstrate the feasibility of using this newly adapted and validated high-content assay to screen chemical or RNAi libraries for the identification of previously uncovered enhancers and suppressors of the apoptotic machinery in live cells. (Journal of Biomolecular Screening 2009:956-969)

Key Words: high-content assay • RNAi HT screening • chemical HT screening • caspase • apoptosis • cancer • live cells

This version was published on September 1, 2009

Journal of Biomolecular Screening, Vol. 14, No. 8, 956-969 (2009)
DOI: 10.1177/1087057109343207
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?